AI Article Synopsis
- Label-free proteomics helps identify disease mechanisms and therapeutic targets, but analyzing ultra-small clinical samples is challenging due to sample loss during pretreatment.
- A new method called MOFs Aided Sample Preparation (MASP) combines several steps of protein analysis using boronic acid-rich metal-organic frameworks, significantly reducing sample size and minimizing losses.
- MASP successfully quantified around 1800 proteins in a small number of cells and identified nearly 3700 proteins in cerebrospinal fluid from stroke patients, proving effective for analyzing precious clinical samples with low protein abundance.
Article Abstract
Label-free proteomics is widely used to identify disease mechanism and potential therapeutic targets. However, deep proteomics with ultratrace clinical specimen remains a major technical challenge due to extensive contact loss during complex sample pretreatment. Here, a hybrid of four boronic acid-rich lanthanide metal-organic frameworks (MOFs) with high protein affinity is introduced to capture proteins in ultratrace samples jointly by nitrogen-boronate complexation, cation-π and ionic interactions. A MOFs Aided Sample Preparation (MASP) workflow that shrinks sample volume and integrates lysis, protein capture, protein digestion and peptide collection steps into a single PCR tube to minimize sample loss caused by non-specific absorption, is proposed further. MASP is validated to quantify ≈1800 proteins in 10 HEK-293T cells. MASP is applied to profile cerebrospinal fluid (CSF) proteome from cerebral stroke and brain damaged patients, and identified ≈3700 proteins in 1 µL CSF. MASP is further demonstrated to detect ≈9600 proteins in as few as 50 µg mouse brain tissues. MASP thus enables deep, scalable, and reproducible proteome on precious clinical samples with low abundant proteins.
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| http://dx.doi.org/10.1002/adma.202401559 | DOI Listing |
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Article Synopsis
- Label-free proteomics helps identify disease mechanisms and therapeutic targets, but analyzing ultra-small clinical samples is challenging due to sample loss during pretreatment.
- A new method called MOFs Aided Sample Preparation (MASP) combines several steps of protein analysis using boronic acid-rich metal-organic frameworks, significantly reducing sample size and minimizing losses.
- MASP successfully quantified around 1800 proteins in a small number of cells and identified nearly 3700 proteins in cerebrospinal fluid from stroke patients, proving effective for analyzing precious clinical samples with low protein abundance.
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